GSM7926245: A549-RIP-seq-HNRNPC-input; Homo sapiens; RIP-Seq
Cells were harvested by RIP lysis buffer, and then the lysates were immunoprecipitated by 5μg anti-HNRNPC antibody. The purified input and HNRNPC-IP RNA samples were then collected for RNA-seq analysis. RIP Sequencing library construction process, firstly, use qubit and 2100 to quantify the RIP fragments and detect the fragment length, then add appropriate Mg2+, heat to interrupt the RNA fragments, add reverse transcriptase, reverse transcription into cDNA, break the RNA strand and form double-stranded DNA by using the broken RNA as the primer and the cDNA as the template, make up the ends of the fragments and add A-tail at the 3' end of the fragment, add Adapter. Add A tail at the 3' end, add Adapter, use 0.8X AMPure beads to remove the excess Adapter, amplify by library PCR, XAMPure beads to remove the excess primer, qPCR to determine the concentration of library, Agilent 2100 to determine the size of the library fragments.